Production of Chloroplast-Derived Human Serum Albumin

Introduction

Human Serum Albumin (HSA) is a monomeric globular preproprotein that accounts for approximately 60% of the protein in serum. HSA is currently the most widely used protein given intravenously and is prescribed in quantities of several grams. HSA is widely used to treat ascites caused by severe burns, hemorrhagic shock, hypoalbuminemia, polycythemia fetus, and liver cirrhosis, and is in great clinical demand. However, it is limited by the only extraction of HSA from blood, that is, the lack of a commercially viable recombinant expression system to prevent the hydrolysis of serum albumin. In addition, HSA derived from human blood has the potential to transmit disease as well as other blood-derived products. Plants are an ideal production system for many recombinant proteins. Scientists have successfully used chloroplast genetic engineering technology as an effective tool for expressing biopharmaceutical proteins in plants, trying to produce recombinant human serum albumin in tobacco leaves and potato tubers.

Fig. 1. Blood sample of patient for albumin test.

Solutions

Our engineers are passionate about chloroplast engineering to produce therapeutic proteins, for research to treat or prevent human or animal disease. Lifeasible is an expert in the field of recombinant protein research and development, with a variety of mature and comprehensive platforms and technologies, providing global customers with professional solutions for the production of chloroplast-derived human serum albumin proteins.

Our engineers attempted to develop a more efficient method of recombinant HSA production. Here, we provide an expression system based on tobacco chloroplast transformation to produce high-purity recombinant HSA. In addition, we developed a cost-effective purification protocol for expressing HSA from potato tubers, transgenic rice seeds. Our solution process is roughly as follows:

(1) HSA cDNA was isolated from a cDNA library constructed with human liver tissue.
(2) The chloroplast transformation vector was constructed by introducing various regulatory elements into the HSA regulatory sequence, that is, inserting the HSA 1.8 kb Eco RI/Not I fragment into the multiple cloning site of the pLD vector.
(3) The vector is delivered into the chloroplast by particle bombardment, and the plant is cultivated for regeneration.
(4) Southern blot analysis whether HSA cDNA was incorporated into the chloroplast genome.
(5) The amount of HSA in chloroplasts of transgenic tobacco was analyzed by ELISA.
(6) Western blot analysis of the expression stability of HSA protein.

Features of Our Solutions

• Enrichment or purification of HSA protein from transgenic plants that are highly susceptible to proteolytic degradation is achieved.
• HSA purification protocols are robust and economical, even at the kilogram scale.
• HSA accumulation increased up to 500-fold in mature plants.
• HSA can form large inclusion bodies and even lead to a significant increase in the size of transgenic chloroplasts.
• Recombinant HSA is biochemically, structurally, functionally and immunologically equivalent to HSA.
• Due to the large amount of HSA used in clinical applications, large-scale production of HSA can be achieved.

The chloroplast-transformed human serum albumin protein is safe and effective, also prevents severe burns, hemorrhagic shock, hypoalbuminemia, polycythemia fetus, and ascites due to cirrhosis, etc. Lifeasible has extensive knowledge and experience in the development and production processes of the chloroplast-derived human serum albumin protein. Our mission is to provide customers with comprehensive, reliable, professional solutions to accelerate your research. If you are interested in our solutions for the production of chloroplast-derived human serum albumin, please contact us at any time.

References

1. Fernández-San Millán A, Mingo-Castel A, et al. (2003) A chloroplast transgenic approach to hyper-express and purify Human Serum Albumin, a protein highly susceptible to proteolytic degradation. Plant Biotechnol J. 1(2):71-79.
2. Ko S M, Yoo B H, Chung H J, et al. (2006) Production of human serum albumin in chloroplast-transformed tobacco plants[J]. Journal of Plant Biotechnology. 33(4): 233.